MANUAL FOR

T.Y.B. PHARM; Sem VI

PHARMACOGNOSY LAB – I
,
SYLLABUS:
EXPERIMENT NO: 1 DATE:

AIM: TO study Compound Microscope and Micrometry

REFERANCE:

1. Dr.K R. Khandelwal, Practical pharmacogonosy, Nirali publications pg no.1.1 to 1.8

2. K. S. Ladhha, herbal drug microscopy ,yucca publication house ,first edition ,march 2003,
page no. 1 to7

MICROSCOPY:
A microscope (micro= small, and scope =to view) is an optical instrument consisting of lens or
combination of lenses. It helps in magnification (enlargement) of the image of an object, which is
too small to be viewed by the naked eye.

There are three principal kinds of microscope: Simple, Dissecting and compound.

SIMPLE MICROSCOPE:

It consists of single lens or magnifying glass, fixed on the suitable frame to view any object e.g. a
hand lens. It is useful where only a low magnification required e.g. for a examination of external
characteristics of crud drugs. Here the magnification obtained is approximately two times.

DISSECTING MICROSCOPE:

This is nothing but the simple microscope with additional features of stage. On which a dissection
can be carried out a suitable hand test for convenience and a mirror to focus the light on the object.
The lens can be raised or lowered by rack and pinion arrangement or moved horizontally for proper
focusing. The magnification in this case is about five times.

COMPOUND MICROSCOPE:

It consists of two sets of lenses. Here one set of lenses of short focal length is used to produce an
enlarge image of an illuminated object at a short distance, which is further enlarged by the second set
of lenses placed appropriately.

A compound microscope consists of the following parts:

The Base: Usually ‘U’ or ‘V’ shaped, which rests on the table. The pillar: an upright bar supporting
the rest of the instrument on an inclination joint.

The stage : A horizontal shelf with graduated mechanical slide holder with X and Y movement for
holding the slide to be examined .The stage bears a hole in the center for transmitting light reflected
up by the mirror.

The mirror: Situated below the stage, reflects light upward through the hole in the stage. The mirror
is usually double faced. The plane face is for initial light intensity and the concave for concentrating
the light on the object.

The Diaphragm: Situated in between the hole on the stage and mirror, regulates the amount of light
reflected by the mirror.

The Body Tube: A cylinder holding a draw tube and moves up and down vertically above the hole
in the stage. The tube is raised or lowered by the coarse adjustment and is use for finding the focus.
The fine adjustment which on being turned produces a very slow motion of the entire framework
which holds the body tube and is used for exact focusing of the higher power lenses.

The ocular or Eye piece: Is to be inserted into the upper end of the draw tube. It consists of 2 plane
convex lenses, the lower and large collective or field lens increasing the field of vision and the upper
and smaller eye lens. Ocular enlarges the image formed by the objective. Midway between the field
lens and eye lens is perforated diaphragm which cut out edge rays from the image, determining the
size of the field of view. Oculars are designated usually with magnification number as 5X, 10X, 15X
etc. or by figures which represent focal length.

The objectives: Are fitted in to the bottom of the body tube or nose piece. Each of these consists of
a system of 2, 3 or more lenses. Objectives, like oculars are usually designated by magnification
numbers as 10, 45 etc. or by focal length. The smaller the number of focal length, the greater is its
magnifying power. If only two objectives accompany your microscope, the lower power objective is
shorter in length. Objective enlarges the object and projects them in the direction of eyepiece.

Magnification and field view: Microscopes are usually fitted with two objectives of 16mm and
4mm, two or three eyepieces and condenser. Different combination of eyepiece and objectives give
different magnification and field of view as indicated in the following table.

Focal length of Approximate magnification with eyepiece (Field of

objective view in bracket )
 Initial 5X 10X 15X
magnifying
power

16mm 10 51(1.65mm) 110(1.1mm) 155(0.89mm)

4mm 45 238(0.42mm) 490(0.25mm) 690(0.21mm)

When using the microscope, it is useful to have the knowledge about the size of the field of view,
e.g. using 4mm objective and 5X eyepiece, field of view is approximately 0.42mm or 420µ.
However, accurate measurements are made with eyepiece micrometer or Camera Lucida. Objectives
are of two types, dry lenses and immersion lenses. The lens is called dry one, if an air space is
present between the tip of objective and the object, if the liquid is present the lens is called an
immersion lens (water immersion lens, if it is water or oil immersion if it is oil). Some microscopes
are fitted with a nosepiece capable of carrying 2, 3 or 4 objectives, which may be adjusted into place
at the lower end of the body tube. Others have a condenser to concentrate the light upon the object to
be examined. When using the condenser use only plan mirror.

How to use Microscope:

 Place the microscope on the table with the arm or pillar nearest to you.
 Turn the lowest power objective into position into position which find light by looking into
ocular (eyepiece) and at the same time turn the mirror at such an angle that it reflects light from the
window or lamp up through the hole in the stage to objective.
 Place the prepared slide in the slide holder on the stage in the horizontal position with the object
right in the center of the hole through which light is reflected from the mirror.
 Make the lower power objective quite close to the slide by turning the adjustment down. Then
while looking through the eyepiece, move the coarse adjustment upward until the object is seen
distinctly. The object if not under the lens may now be brought into the field by moving the X-Y
movement very slowly while looking through the eyepiece. Then slowly turn the fine adjustment to
improve the focus.
 Regulate the quantity of light with the iris diaphragm to improve the clarity of the field.
 To focus with the light power objective, first find the object with the low power and arrange in
centre of the field. Raise the low power objective by means of coarse adjustment. Then turn the high
power objective into position and lower until the objective front lens nearly touches the cover glass.
A slight movement of the fine adjustment should show the object clearly. Never try to focus with
the high power objective while looking through the eyepiece because of the danger of cover glass
and damaging the tissue lying underneath. It may also cause damage to the delicately mounted
lenses.

Care of Microscope:
 While carrying the microscope from one place to another, hold it firmly by the arm in an erect
position so that the ocular which is fitted loosely in the drawtube may not fall out.
 Do not allow the dry object or the liquid in the draw tube, may not fall out.
 Do not touch the ocular or stand with cleaning cloth soaked by reagents.
 Raise the body tube sufficiently while changing from low power to high power objective to avoid
damage to the objective or the mounts.
 If the lens is soiled it may be cleaned with clean cloth wetted with few drops of Xylol.
 Never observe objects without putting a cover glass.
 For removing slide from the stage, first raise the body tube and slowly slide it out of stage.
 The slide must be prepared in such a manner that the stage is never wetted with any solvent or
reagent.
 When not in use, microscope must be kept covered.
Histological techniques:
For obtaining satisfactory result of the following items are needed:
1. A sharp shaving blade for sectioning
2. A scalpel
3. Dissecting needle
4. Forceps
5. Watch glasses
6. A sufficient number of Microscopic slide
7. Cover slips
8. A 0 no. Camel hair brush
9. A piece of clean cloth
10. A record book
11. Pencil, pen etc.

The preparation of plant material from microscopic examination and the reagents used are described
below. Preparation of material for section cutting:

The drug available for pharmacognostic study are generally in the dry condition. The necessity tales
soaking of these drugs to soften them sufficiently, which permit easy sectioning of drug. The
duration of soaking depends up on nature of tissue of drug.

Drug Material Soaking time

Clove, Coriender, Digitalis, Ephedra, Fennal, Senna 24hrs. in water

Cardamom, Cassia, Cinchina, Kurchi 2-5 days in chloroform water

Qussia, Quillaia, Glycyrrhiza ,Nuxvomica, 2-5 days in chloroform water

Rauwolfia

Chloroform water prevents microbial growth. Drugs which are available in fresh form like Datura,
Eucalyptus, Ginger, Neem , Vasaka etc. should be kept in water to avoid cell shrinkage by loss of
water.
SECTIONING:

The material to be sectioned is held between the thumb and for finger of the hand. Using a sharp
razor held in right hand, thin sections are made by drawing the razor across the object in quick
succession, sliding over the forefinger with the edge of forefinger pointing towards you. Transfer the
section cut, into water kept in watch glass with the help of Camel’s hair brush. In case of tender and
flexible material such as fresh leaf, the section can be taken conveniently by placing it between the
two flat surfaces of pith. A pith is usually a piece of potato( about 3×1×1cm ) in which a
longitudinal slit of two cm deep is made into which the material to be sectioned is placed is placed
and the section are taken as described above.

Three types of section are possible:

1. Tranverse section (TS): Is made in horizontal place at right angle to the long axis of
material.
2. Radial- longitudinal section (RLS): is made in a longitudinal plane parallel to the long axis
of the material along its radius.
3. Tangential-longitudinal section (TLS): is made in longitudinal plane parallel to the long
axis of the material and to the tangent.

In addition to the above three a surface view can be obtained by pealing of outer most layer e.g.
epidermal area of the leaves or the cork layer of the bark.

Techniques Of mounting:

From the watch glass containing the section transfer section to the center of glass slide, put 2-3 drops
of chloral hydrate solution on it, heat the slide very gently by passing it to and fro over low flame
when bubble start to appear stop heating, add a drop of glycerin- water solution to avoid drying of
the preparation and crystallization of chloral hydrate and place the cover glass carefully. To avoid air
bubbles getting entrapped, the cover glass is allowed to stand on one side and gently squeeze the
liquid between the cover glass and slide. Chloral hydrate is good clearing agent, which dissolves
starch, proteins, chlorophyll, resins and volatile oils which causes expansion of shrunken cells. The
method describe above is useful for observation of calcium oxalate crystals, as chloral hydrate does
not dissolve calcium oxalate.

For staining the preparation after treatment with chloral hydrate, add a drop of phloroglucinol
reagent on the section, allow it to evaporate and then add then add a drop of hydrochloric acid and
wait for 2 min. Then add then mounted in water. Their presence can be conforming by staining with
a iodine solution. The powder of crude drugs can also be mounted describe above for identification
of various characteristics.
Reagent Used:

Chloral hydrate solution: Dissolve 25gm of chloral hydrate in 10ml water.Glycerin- water solution:
Prepare by adding 1:1 glycerin and distilled water. Iodine water: Add as much iodine to distilled
water as it well dissolves.Phloroglucinol solution: Dissolve 1 gm of Phloroglucinol in 50 ml of 95%
alcohol. Use this solution within.Conc. Hydrochloric acid.

Fig 1: EYEPIECE MICROMETER Fig 2 : STAGE MICROMETER

Fig 3: 51 divisions of eyepiece micrometer coincide with 20 division of stage micrometer

MICROMETRY:

Micrometry is the determination of particles with the help of microscope. The accessories required
for the of measurement are eyepiece micrometer and stage micrometer.
 I. Stage Micrometer: It is a slide .It contains a standard scale length of 1mm which is divided into
100 divisions.

1division of stage micrometer = = 0.01m

or

1division of stage micrometer = =10micro

II. Eyepiece Micrometer: It is a circle of glass with scale attached on the surface. It is suitable for
insertion inside the ocular and used during the operation of measurement. A convenient form is a
linear scale of 1mm divided into 100 divisions.

CALIBRATION OF ONE DIVISION OF EYEPIECE MICROMETER

a. Put the eyepiece micrometer on the diaphragm of the ocular

b. Put the stage micrometer on the stage of microscope
c. See the lines of two micrometer coincide. Count the number of lines required to coincide.

CALCULATION:
1division of stage micrometer = 0.01mm = 10µ
If, 6division of eyepiece micrometer = 4 division of stage micrometer.
The value of one division of eyepiece micrometer is calculated as follows-
1division of stage = 0.01mm=10µ
4division of stage =40micron
Now, 6 division of eyepiece micrometer = 4 division of stage
=40µ

1division of eyepiece micrometer = =6.67µ

Now, to measure any object under consideration, place the prepared slide on the stage. See how
many lines are required.

If 6 lines (divisions) are required then, 6×6.667µ=40.002

 MICROSCOPIC DRAWINGS

Swift Ives Camera Lucida and Abbe Model are the commonly used instrument to trace
the image of an object under microscope on the paper. The image of object under the microscope
can be traced on paper with the help of swift- Ives camera lucida and a drawing board , whole
inclination can be adjusted . The camera lucida fixed over the eyepiece of the microscope. Figure
shows path of light from the object passing directly to the observer's eye through an opening in the
silvered surface of the left hand prism. At the same time light from the drawing paper and pencil is
reflected by the right hand prism and by the silvered surface, so the pencil appears superimposed on
the object enabling it to be traced on the paper. Other type of camera lucida working on same
principle is also available. The abbe drawing apparatus is another form of apparatus, which can be
used to trace the image of an object without any inclination in the board. It utilizes a plane mirror
carried on a side arm, instead of the adjustable prism, with the mirror at 450 to the bench surface.

Camera lucida or drawing ocular is useful for tracing a magnified image of the object under
microscopical studies with proper adjustments of camera lucida and illumination. It is possible to see
simultaneously the drawing paper, the pencil point and the object under microscope and it is then
easy to trace the require cutlines. This is much quicker and more accurate than the most skilled free
hand drawing, but it requires the subsequent addition of details by free hand.

Swift Ives, camera lucida and abbe model are commonly used instruments. Swift Ives, camera
lucida consist of a prism fitted over the microscope should be in a vertical position and microscope.
Lamp should be carefully adjusted, so that illumination on the drawing paper, placed at the site of
the microscope is equal to that on the mounted preparation. The drawing pare should be supported
drawing board and if necessary tilt it at correct angle to avoid distortion. Test the equal illumination
on all sides of drawing board. By placing a stage micrometer on stage of the microscope and tracing
its division by co-inciding the image of tip of pencil and graduation of stage micrometer. If division
drawn are not equal, then the angle of tilted board is readjusted, until equality is obtained. Remove
the stage micrometer and place a mounted slide with specific drug preparation on stage of
microscope, after drawing of stage micrometer scale on drawing sheet, the distance between 2 point
is measured with the help of scale. For measurement of object, replace the stage micrometer with
object under study without disturbing the adjustment. Trace the outline of object same as that of
stage micrometer. Co-incide the tip of pencil in margin of object finish the drawing. Measure the
dimension of object. By this arrangement, the circle over the eyepiece comes just above the cornea
of eye and camera lucida. The field of view is not in anyway reduced and changed. All that can be
seen directly through the eyepiece perfectly disciplined camera lucida, while the drawing being
viewed directly on drawing sheet but not drawn on field.

In practice the drawing board should be adjusted at particular angle approx 45 0 and the image
accurately focused without the eyepiece. The camera is slide on eyepiece and pushed down more or
less until the microscopical image is seen directly and illumination of the field is equal throughout.
The drawing paper is placed on the table, immediately under the camera. The observer will then see
the microscopical image projected on paper. At the same time waving the pencil point directly the
whole pupil of eyepiece is available for both images (object and drawing board) the diaphragm on
the apparatus being considerably larger than the pupil so it may be necessary to balance the
illumination either by subdividing the light in the microscope or by increasing it on drawing paper. It
will generally be found that when the object is ion luminious field the light on the object may be
advantageously subdivided by glass around it or similar measures. The eye may be removed as
required from the camera to minimize the parallax produced because of camera lucida and change in
intensity of illumination.The drawing paper should be kept at a distance of distinct vision according
to change in camera lucida and the person who is studying that object.

a. Swift Ives Camera Lucida:

The image of an object under the microscope can be traced on paper with the help of Swift Ives
Camera Lucida. The Camera Lucida fixed over the eyepiece of the microscope.
The Swift Ives Camera Lucida consist of prism fitted over the microscope eyepiece. While using
Camera Lucida, the microscope should be in vertical position and microscope lamp should be
carefully adjusted so that the illumination on the drawing paper placed at the side of microscope is
equal to that on the mounted preparation.

b. Abbe Model:
The Abbe drawing apparatus is another form of apparatus, which can be used to trace the image of
an object without any inclination in the board. It utilizes a place mirror carried on the side arm,
instead of adjustable prism, with the mirror at 45˚ to the bence surface.
EXPERIMENT NO: 2 DATE:

AIM: Identification of Starch grains by Microscopic Evaluation.

REFERENCE:

1 .K.S.Laddha,herbal drug microscopy,yucca publishing house, pg no. 17 &18

2. DR. K.R. Khandelwal,practical pharmacogonosy,nirali publication,pg no. 21.1-21.2

STARCH.
a. BOTENICAL SOURCE-

Starch consists of polysaccharides granuales separated from mature grains of

corn(Zea mays), wheat( Tritum aestium), rice(Oryza sativa) all three belonging to
family. Gramineae, or from tubers of potato(solanum tuberosum) Fam. Solanaceae.
Starch occurs as irregular, angular, white masses or fibers with no odour and slightly
mucilaginous taste. The hilum, which appears as a darker or lighter point is the
starting point for the formation of granules in the leucoplast. The hilum may be central
or may be nearer to the one end i.e eccentric. Fissure originating from hilum can be
seen in variably. The stratinous or concentric rings are fine lines surrounding the
hilum, which are formed by deposition of successive layers of starch around the
hilum.
Corn starch.
Polygonal,rounded granules up to 35µ in diameter, hilum distrinct, central ,triangular
or 2to5 radiate, striations absent
Wheat starch.
Simple, lenticular, which are rounded or oval 2-45µ in diameter, hilum acentral
point;striations,faintly marked and concentric

Potato starch.
Simple granules, irregularly ovoid or spherical and sub spherical 30-100µ in diameter;
hilum-eccentric; striations well marked and concentric.

Rice starch.
Compound granules (aggregation of large number of simple granules)up to 150
component granules,2 to 10µ,polyhedral with sharp, hilum minute central point, rarely
conticuous ,striations absent

b. CONSTITUENTS:

Starch contains two complex poilysaccharides, amylopectin(α-amylase)and

amylose(β-amylase) usally in the proportion of 1:2 amylase is soluble in water and
gives a bright blue colour with iodine solution while amylopectin is insoluble in water
but swells in it producing a viscous paste when boiled. amylopectin gives bluish –
black colour with iodine solution

c. STARCH USES:

Starch is used as pharmaceutical aid particularly in tablets as disintegrating agent,

lubricant, filler and binder. It is widely used in dusting powders because of its
absorbed properties. Starch is also used for the manufacture of liquid glucose,
dextrose and dextrin

d. SUBSTITUENTS:

Tapioca starch or cassava starch from rhizomes of Manihot utilissiama and Manihot
aipi, fam Euphortbiaceae.
Sago starch from metroxylan sagu,M.rumphii ,Fam palmae.
Sweet potato starch
Brazilian arrowroot starch

RESULT:
EXPERIMENT NO: 3 DATE:

AIM: To study calcium oxalate crystals in given sample

REFERENCE:
Dr. K.r. khandelwal, practical pharmacogonosy,nirali publication,pg no. 9.1-9.2

CALCIUM OXALATE CRYSTALS

Crystals of calcium oxalate occurs in many plants. This are form by reaction of
calcium salt (which have been absorbed from the soil) with oxalic acid(by product of
protein and another metabolic process). Among the various cell content, calcium
oxalate crystals of different type are found in different organs of the plant. They may
be present in almost all parts of plant

Calcium Oxalate crystals occurs in two forms:

1) Trihydrates - belonging to tetragonal system

2) Monohydrate- belonging to monoclonal system

Crystals belonging to the tetragonal system have three axis at right angle to each other,
two of these axis are equal in length, the third being of different length.
Crystals belonging to the monoclinic system have three axis of unequal length, two of
which are obliquely inclined to each other and other two are right angles to these two
Usually Calcium Oxalate crystals are described according to their general form and size.

1] Solitery(single) crystals- Usually in the form of prism, occurs as sharp angular bodies.eg:
Quassia, Senna.
2] Twin Crystals- Two prisms are united.eg: Hyoscyamus.
3] Rosette Aggregates- It consists of numorous small prism appears like rosette or star.eg: Rhuhab,
Senna.
4] Columnar Crystals- Elongated prisms.eg: Quillaia Saponaria.
5] Acicular or needle shape crystals- Single needle shape crystals usually found scattered in
parenchymatous cells of cinnamon bark etc.
6] Microcrystals (sandy crystals or microsphenoidals)- these are arrow-shaped minute completely
filling the cells in which they are occur.eg: Belludonna, Tobacco etc.
7] Crystals Fibers - These are superimposed parenchyma cells each of which contains single prism
or a rosette aggregate.

Significance:
-They give protection to the plant against birds and animals.
-They have great diagnostic value.
-Presence of or absence of crystals useful in identification of crude drugs.
-Helps in identification of adultrants.

RESULT:
EXPERIMENT NO: 4 DATE:

AIM: Identification and Trichomes by Microscopic Examination

REFERENCE:

Dr.KR. Khandelwal,practical pharmacognosy,nirali publication pg no.11.2-11.4

TRICHOMES (EPIDERMAL HAIRS):

Trichomes are more elongated outgrowth of one or more epidermal cells , and consists of two parts,
a foot and root embedded in the epidermis and a free projecting portion termed as body. Trichomes
usually occur in leaves but are also found to be present on some other parts of the plant.eg; kurchi
bark. Trichomes are rarely present on the leaves of beaeberry ,buchu,henna etc.

Functions of trichomes: Trichomes or hairs are adapted to many different purposes. A dense
covering of Trichomes prevents the damage by insects and the clogging of stomata due to
accumulation of dust. Trichomes perform the function of secreting volatile oil.

Types of Trichomes:

1) Covering trichomes or clothing hairs or non glandular.

2) Glandular trichomes.

A) Covering Trichomes
a) Unicellular Trichomes

1.Linear, Strongly waved , thick walled trichomes –yerba santa

2. linear, thick walled and wanty trichomes-Damiana
3. short conical wanty trichomes-senna
4. large , conical, , longitudinally striated trichomes-lobelia
5. long,tubular,fluttened and twisted trichomes-cotton
7.lignified trichomes- nux vomic, strophanthus
8. short, sharp, pointed, curved, conical, trichomes-cannabis
9. unicellular stellate trichomes- deutezia scabra

b) Multicellur unbranched Trichomes.

1. Uniseriate, bicellular, conical-Dhatura
2. Uniserita ,3-4 celled long – Stramonium belladonna
2. Biserita – calendula officinalis
3. Multiserita – male fern

c) Multicellular branched trichomes

1.stellate-hamamelis, helicters isora fruit, kamala

2.paltate ( shield-like structure)cascarilla
3.Candelebra ( branched)- Egyptian henbane, rosemary, uerbascum thapus
4. T-shaped trichomes – Artemisia, pyrethrum

B) Glandular Trichomes

a) Unicellular glandular trichomes

without stalk termed as sessile trichomes-piper betel and vasaka
b) Multicellular glandular trichomes
1. Unicellular stalk with single spherical secreting cell at the apex
–Digitalis purpurea
2. Uniserita multicellular stalk with single spherical cell at the apex
-Digitalis thapsi.
3. uniseriate stalk and biseriate secreting head
– santonica and other plant of family composiate
4. Multicellular , unicseriate stalk and multicellular head –hyoscyamus
5. Short, unicellular stalk and head fromed by a rosette of 2-8 club shaped cell – Androgrophis
RESULT-
EXPERIMENT NO: 5 DATE:

AIM: To study stomata in given sample

REFERENCE:

1. K.S. Laddha,Herbal drug microscopy,yucca publication,pg no.10-11

2. DR K.R.khandelwal,practical phamacognosy,nirali publication,pg no.11.2

THEORY:

Stomata are minute openings usually found in the epidermis of the leave as in digitalis,senna etc. or
in young green stems as in ephedra, in flower as in clove and in fruit as in fennel , orange as orange
peel. This opening are surrounded with a pair of kidney shaped cells called guard cells. The term
“STOMA” is often applied to the stomatal apparatus which consist of slit like opening along with the
guard cells.The epidermal cells surrounding the guard cells are called neighbouring cells or
subsidiary cells.This in many cases as in digitalis etc. resemble the other epidermal cells, but in large
number of plants they differ in size, arrangement and shape from the other epidermal cells. On the
basis of the characteristic of the guard cells and subsidiary cells, Stamatas can be classified as:

 CARYOPHYLLACEOUS OR DIACYTIC (cross-celled):

The stoma is accompanied by two subsidiary cells,the long axis of which are at right angles to that of
the stoma.This type of stoma is also called as the LABIATAE type as it is found in many plants of
the family labiatae such as spearmint,peppermint,thyme,tulsi and vasaka.
 CRUCIFEROUS OR ANIISOCYTIC (unequal-celled):
The stoma is surrounded by usually three subsidiary cells of which one is markedly smaller than the
others.This type of stoma is called the SOLANACEOUS type as it is found in many plants of the
family solanaceae
Such as belladonna,datura,hyoscyamus,stramonium,tobacco.It is also found in many plants of the
family compositae.
 RANUNCULACEOUS OR ANOMOCYTIC (irregular-celled):
The stoma is surrounded by a varying number of cells in no way differing from those of the
epidermal cells as in buchu, digitalis,eucalyptus,heena, lobelia, neem.
 RUBIACEOUS OR PARACYTIC(parallel-celled):
This stoma is surrounded usually by two subsisdary cells, the long axis of which are parallel to that
of stoma as in senna and many rubiaceous plants.
 ACTINOCYTIC (radiate-celled):
The stoma is surrounde by circle of radiating cells,as in Uva ursi.stomata with dumbelled shaped
guard cells and crescent shaped subsidiary cells on either side occurs frequently in longitudinal rows
altering with long cells as in grasses.

FUNCTIONS AND DISTRIBUTIONS OF STAMATA

Stomata perform the function of gaseous exchange and transpiration in the plants body. They are
most abundant in the lower epidermis of a dorsiventral leaf and less abundant on the upper in
isobilateral leaf, stomata remain confined to the upper epidermis alone, in submerged leaves no
stoma is present.In buchu and neem ,stomata are present only on the lower surface, while in case of
bellodona,datura,senna etc. stomata are present on both the surface.
The distribution of stoma shows great variation between upper and lower epidermis. In desert plants
and in those showing xerophytic adaptations eg.ephedra, agave, oleander etc.stomata are situated in
grooves or pits in the stem or leaves. This is special adaptation to reduce excessive evaporation as
the stoma sunken in the pits are protected from gusts of wind.

PROCEDURE:

1. Take a small amount of powder of a given sample in a test tube, add chlorhydrate and mix
well.
2. Take the mixture on watch glass and mix properly again.
3. Take this mixture on a slide add a drop of water with the help of brush.
4. Put the coverslip without any air bubble.
5. Observe under microscope at 10x and 45x respectively.
RESULT:
EXPERIMENT NO: 6 DATE:

AIM: To isolate starch from potatoes.

REFERENCE:

1. Practical Pharmacognosy, C.K. Kokate, 5th edition, Vallabh Prakashan, pg. no. 159,160.

MATERIALS AND REAGENTS:

Waring blender, centrifuge, stirrer, shaking sieves, oven, distilled water, potatoes.

PROCEDURE:

1. Wash potatoes thoroughly with water to remove adhering soil and earthly matter and reduce
to fine slurry with water in a blender.
2. Pass the slurry through shaking sieves in order to remove the cell debris and other impurities.
3. Allow the milky liquid to settle down. Decant the supernatant liquid. Wash starch 2-3 times
with distilled water with constant stirring.
4. Centrifuge the milky liquid, dry it in oven at a low temperature and powder.

The yield of starch is approximately 10 percent and it gives blue colour with weak iodine solution.
RESULT
EXPERIMENT NO: 7 DATE:

AIM: To determine alcohol extractive value of Ginger powder.

REFERENCE:

1.Practical Pharmacognosy by Dr.K.H.Khandelwal ,pg no. 23.10

2. Pharmacognosy by C.K.Kokate ,Gokhale and Purohit ,pg no . 112(6.21)

THEORY:

EXTRACTIVE VALUES

 Useful for the evaluation of crude drug

 Give idea about the nature of the chemical constituent present in the drug
 Useful for the estimation of constituents extracted with the solvents used for extraction
 Employed for material for which as yet no suitable chemical or biological assay exist

ALCOHOL SOLUBLE EXTRACTIVES:

Alcohol is an ideal solvent for extractive of various chemicals like tannins, resin etc. Therefore this
method is frequently employed to determine the approx resin content of drug. It is also used as an
official method for assay in case of myrrh and asafetida .Generally, 95% ethyl alcohol is used for
determination of alcohol soluble extractive. In some diluted alcohol may also be used, depending
upon solubility of the constituent of crude drug.

PROCEDURE:
Weigh about 4 gm of the coarsely powdered drug in a weighing bottle and transfer in to a dry 250 ml
conical flask.

Fill a 100 ml graduated flask to the delivery mark with the solvent (90% alcohol). Wash out the
weighing bottle and pour the washing with the remainder of the solvent into the conical flask.

Cork the flask and set aside for 24 hours shaking frequently (maceration)

Filter into 50 ml cylinder. When sufficient filtrate has collected, transfer 25 ml of the filtrate to a
weighed thin porcelain dish, as used for the ash values determination

Evaporate to dryness on a water bath and complete the drying in an oven at 105 0 C for 6 hours

CALCULATION:

Wt. of petri dish = a =………………………

Wt. of petri dish + extract = b =……………………………..

Wt. of extract = b-a=………………………………. = …………………….

------g of drug = ------ ml of ------% alcohol

------g of alcohol =X g of extract

X = ------ x ------

= -------%

Alcoholic extractive value of ginger was found to be --------%

For Liquorice ,
Wt. of Petri dish =

Wt. of extract petri dish =

Wt. of extract =

------g of drug = ------ml of ------%

------g of alcohol = Xg of extract

X= ------ x ------

------

Alcoholic extractive value of Liquorice was found to be -------

RESULT:

From the above observation the alcoholic extractive value Ginger and Liquorice powder was found
to be -------- % w/w and -------- % w/w respectively.
EXPERIMENT NO: 8 DATE:

AIM : To determine Water extractive value of Ginger and Liquorice powder.

REFERENCE:

1. Practical Pharmacognosy by Dr.K.R.Khandelwal,pg no. 23.10

2. Pharmacognosy by C.K.Kokate, Ghokhale and Purohit .

THEORY:

Extractive value:

 Useful for evaluation of a crude drug.

 Gives an idea about nature of the chemical constituents present in a crude drug.
 Useful for estimation of constituents extracted with the solvent used for extraction
 Employed for material for which as yet no suitable chemical or biological assay exists.

Water soluble extractives:

This method is applied to drug which contain water soluble active constituents of crude drugs, such
as tannins, sugar, plant acids, mucilage, and glycosides.
PROCEDURE:

Weigh about 4 gm of the coarsely powdered drug in a weighing bottle and transfer in to a dry 250 ml
conical flask.

Fill a 100 ml graduated flask to the delivery mark with the solvent (chloroform water).
Chloroform acts as preservative. Wash out the weighing bottle and pour the washing with the
remainder of the solvent into the conical flask.

Cork the flask and set aside for 24 hours shaking frequently (maceration)

Filter into 50 ml cylinder. When sufficient filtrate has collected, transfer 25 ml of the filtrate to a
weighed thin porcelain dish.

Evaporate to dryness on a water bath and complete the drying in an oven at 105 0 C for 6 hours

CALCULATION:

Wt. of empty petridish =______g

Wt. of petridish = ______g

Wt. of extract = ______ - ______

= ______g

______g of drug = ______ml of ______% water chloroform (water : CHCl3 )

______ of water soluble extract = ______g

______ml of water soluble extract =Xg

X =------ x ------

_____

X = ______% w/w

Water soluble extract value of given drug (Ginger ) = ______%

Water soluble extract value of Liquorice =

______g of drug = ______ml [water –CHCl3 (90:10)]

______ml = ______g

______ml = y

y = ------ x ------

_____

y = ______%w/w

Water soluble extractive value of given drug Liquorice = ______%w/w

RESULT:

From above observation, the water soluble extractive value of Ginger and Liquorice was fond to be
______% w/w and ______% w/w respectively.
EXPERIMENT NO: 9 DATE:

AIM: Determination of stomatal nomber and stomatal index.

REFERENCE:

1. Herbal drug microscopy by T.N vasudevan and K.S ulka publishing house pg no 20
2. Practical pharmacognosy Techniques and experiments by Dr. K.R. khandelwal, nirali
publication pg no 24.1 to 24,2.

THEORY –

Stomatal no – stomatal number is the average no of stomata per sq.mm of the epidermis.It is more
significant in evaluation of leaf drug.

Stomatal index – stomal index is the percentage of epidermal cell of leaf which have been
converted into stomata.

T= S * 100

T+S

Where S = no of stomata/unit area

E = no of epidermal cells in the same unit area

STANDARD VALUE

A. Stomatal number

LEAF NO OF STOMATA PER SQ.MM

UPPER SURFACE LOWER SURFACE

Datura stramonium 59 to 83 to 140 145 to 200 to 254

Cassia angustifdia 180 to 200 to 273 189 to 220 to 257

Atropabellodona 7.5 to 10 to 17.5 77.5 to 138 to 176

Cassia auriculata 100 to 200 130 to 260

B. Stomatal index :

Atropa belladonna lower surface –19.5 to 21.6 to 23.9

Cassia aculifolia both surface –16.7 to 17.6 to 18.8
Crusia angustifolin both surface –17.1 to 18.7 to 20.0
Erythroxylum lower surface –12.0 to 13.3 to 15.4

PROCEDURE:

Stomatal number

1. Clear the piece of the leaf by boiling with chloral hydrate solutionor alternatively with
chlorinated soda , peel out upper and lower epidermis. Separetly by means of forceps keep it on slide
and mount in glycerin water.
2. Arrange the camera lucida and drawing board for making the drawing to scale.
3. Draw a square of 1 mm by means of stage micrometer.
4. Place the slide with clean leaf ( epidermis ) on the stage trace, the epidermal cell and stomata
on the paper.
5. Count the no of stomata present in the area of 1 sq.mm include the cell if atleast half of its
area ;ies within the square
6. Record the result for each of ten fields and calculate the average number of stomata/ sq.mm

Stomatal index :
Step no 1, 2, 3, 4 are similar as mentioned in the determination of stomatal no.
7. Count the no of stomata, also the number of epidermal cells in each field.
8. Calculate the stomatal index using the above formula.
9. Determine the value of upper and lower surface ( epidermis ) separetly, for determination of
average index , not less than 10

RESULT :
EXPERIMENT NO: 10 DATE:

AIM: To determine vein-islet and vein termination number of senna leaflet.

REFERENCE:

1. Practical pharmacognosy techniques and experiments by K. R. Khandelwal and DR. vrunda

Sethi, Nirali prakashan, pg no. 24.3 ,24.4.
2. Harble Drug Microscopy , Edited by T. N. Vasudevan, K.S. Laddha, Yucca publishing house,
pg no. 20

THEORY:

Vein islate no is the no of vein islates per sq.mm .

vein islate is the area of photosynthetic tissue and circulated by ultimate division of the conducting
strands.ion

vein termination NO:- It is the no of vein termination per sq.mm of leaf structure.

Significance

Vein islet and vein termination no. can be used for the identification of plants and can be used as a
tool for standardistion of crude drug to prevent the use of an adultrated drug.
PROCEDURE:

 Take senna leaflet and to a leaf prepare a specimen by cutting the leaf from midrib to margin
( near about 2-4mm2 ).
 Take these pieces in a test tube containing a porcelain piece.
 Add chloral hydrate (1ml) and few drops of glycerin water, attach cotton plug wetted with
water to the test tube. Heat the test tube till bubble appears.
 Then remove out the chloral hydrate with tap water.
 Repeat this treatment 2-3 times till the specimen turns colorless (Pale yellow).
 Keep it on slide and mount with glycerin water.
 Arrange a camera lucida and drawing board for making the drawing to scale.
 Draw the scale with the help of stage micrometer.
 Trace out the veins which are seen from the eyepiece, completing the outlines of those islet .
 Select the particular area ( square or rectangle) in a drawing and count the number of vein
islets and vein termination within that area.
 Then calculate vein islet number and vein termination number in 1sq.mm area.

RESULT:

 Vein islet number of senna leaflet was found to be ------- per square mm area.
 Vein termination number of senna islet was found to be ------ per square mm area.
EXPERIMENT NO: 11 DATE:

AIM: To determine the palisade ratio of senna leaflet.

REFERANCE: Dr.K.R.Khandelwal, Dr. Vrunda Sethi, Practical Pharmacognosy Techniques and

Experiments, Nirali Prakashan, Pg no. 24.2 and 24.3

THEORY:
PROCEDURE:

1. Clean the piece of the senna by boiling in a chloral hydrate solution.

2. Arrange the Camera Lucida and drawing board for making drawings
3. Using 4mm objectives trace of the outlines of four cells of epidermis on black sheet.
4. Then focus on Palisade layer and trace of sufficient cells to cover the tracing of the epidermal
cells. Complete the outlines of those Palisade cells which are intersected by the epidermal cells.
5. Count the Palisade cells under the four epidermal cells.(include the palisade cells in count
when 50% portion of the palisade cells in the epidermal cells)
6. Calculate the average number of cells beneath single epidermal cells this figure is palisade
ratio
7. Repeat the determination for five groups of four epidermal cells from different parts of the
leaf. Take the average of the results for five groups. These average is the palisade ratio of the leaf.

RESULT: The Palisade ratio of senna leaflet was found to be

EXPERIMENT NO: 12 DATE:

AIM: To determine number of stone cells in Cinnamon powder by lycopodium spore method.

REFERENCE:

1. Practical pharmacognosy by Dr. K. R. Khandelwal pg. no 23.4

2. Textbook Pharmacognosy Gokhale, Kokate, Purohit, pg . no
3. Textbook by Wallis pg. no 74

REQUIREMENTS: Lycopodim spore, cinnamon powder, glycerine, phloroglucinol HCL (1:1)

THEORY:

LYCOPODIUM SPORE METHOD

It is an important analytical technique for powdered drugs, especially when chemical and other
methods of evaluations of crude drugs fails as accurate measures of quality. It is inexpensive with
official status. Lycopodium spores are very characteristic in shape and appearance and exceptionally
uniform in size (25µm) on an average 94,000 spores per mg of powdered lycopodium are present.

A powdered drug is evaluated by this technique if it contains

1. Well defined particles which may be counted eg.: starch grains or pollen grains.
2. Single layered cells or tissues, the area of which may be traced under suitable magnification
and actual area calculated or
3. The object of uniform thickness, the length of which can be measured under suitable
magnification and actual area calculated.

The percentage purity of an authentic powdered ginger is calculated using the following equation,

N x W x 94,000 x 100/S x M x P = % purity of drug

Where,

N = number of characteristic structure (eg. Starch grains) in 25 fields

W = weight in mg lycopodium taken

S = number of lycopodium spores in the same field

M = weight in mg of the sample, calculated on the basis of sample dried at 1050C

P = 2,86,000 in case of ginger starch grain powder.

Lycopodium spore method can be used for evaluation of powdered kurchi,clove, ginger, cardamom,
nutmeg, umbeliferous fruits, etc.

STONE CELLS

Scleroids, stone cells or sclerenchymatous occurs in the parenchyma of many barks. These cells are
parenchymatous elements and may be rounded, polyhedral or prismatic, they have lignified walls
and the lumen may vary from a narrow, branching, slit like hollow to a fairly large, sub-rectangular
cavity. The walls commonly show striations and are performed by tubular pits, which are often
branched. The external opening of the pits appear as small, circular or irregular pores dotted over the
surface view. The value of these cells for diagnostic purposes is illustrated by the fact that they are
absent from frangula bark, but are present in the very similar cascara bark, they are few in quillaia
and very numerous in cinnamon and cassia.

PROCEDURE:

1. Dry the powdered drug at 1050C and determine its steady wt at RT

2. Weigh accurately powdered material and lycopodium spores and mix them in proportion of
1:1 powdered drug to lycopodium has been found to be satisfactory. Mix them on a glass plate with a
flexible spatula.
3. Make a smooth paste by adding suspending fluid (oil or glycerin : water, 2:2)
4. Add a staining agent as Phloroglucinol : HCL reagent.
5. Keep the solution for few sec for getting stained properly and then add a drop of solution on
glass slide and cover it with cover slip and observe under microscope (45X)
6. Calculate the lycopodium spores as well as stone cells in 25 fields.

OBSERVATION:

Table no. 1

Field. 1 2 3 4 5

Lycopodium spore

Stone cell

Table no. 2

Field. 1 2 3 4 5

Lycopodium spore

Stone cell

Table no. 3

Field. 1 2 3 4 5

Lycopodium spore

Stone cell

Table no. 4

Field. 1 2 3 4 5

Lycopodium spore
 Stone cell

Table no.5

Sr. no. 1 2 3 4 5

Lycopodium spore

Stone cell

CALCULATION
Total number of lycopodium spores in 25 fields = x=
Total number of stone cells in 25 fields = y =
If _x_ no of lycopodium spores have _y_ no of stone cells in 25 fields.

Therefore,
Percentage of number of stone cells = ______ x 100
in given sample

= ________ %

RESULT:
From the above observation, percentage of number of stone cells in given sample of kurchi powder
was found to be _______ %
EXPERIMENT NO: 13 DATE:

AIM : To determine Total Ash value of given crude drug powder.

REFERENCE :

1.C.K.Kokate,Practical pharmacognosy, fifth edition , vallabh prakashan, page no. 123,124.

2.Biren Shah, A.K.Seth , Textbook of pharmacognosy and phytochemistry , second edition , page no.
119.

THEORY :

Principle: The ash content of a crude drug is generally taken to be the residue remaining after
incineration like carbonates, phosphates and silicates of sodium, potassium, calcium and magnesium
etc. It usually represents the inorganic salts naturally occurring in the drug and adhering to it, but it
may also include inorganic matter added for the purpose of adulteration so for the determination
different types of ash value are used in detection of crude drugs like total ash , acid-insoluble ash,
water soluble ash and sulphate ash . And this standards have been established for a number of
official drugs.

Significance: Ash values are helpful in determining the quality and purity of crude drugs, especially
in powder form. And it is also useful for detecting low- grade product, exhausted drug, and excess of
sandy or earthy matter.

TOTAL ASH VALUE:

Defination :
The total ash is the residue remaining after incineration. The acid insoluble ash is the part of the total
ash which is insoluble in diluted hydrochloric acid.
Procedure :
Incinerate about 2 to 3 g accurately weighed, of the ground drug in An tared platinum or silica dish
at a temperature not exceeding 450º until free from carbon, cool and weigh. If a carbon free ash
cannot be obtained in this way, exhaust the charred mass with hot water, collect the residue on an
ashless filter paper, incinerate the residue and filter paper, add the filtrate, evaporate to dryness, and
ignite at a temperature not exceeding 450º. Calculate the percentage of ash with reference to the air-
dried drug.

Significance :
Total ash value is useful in detecting the crude drugs that are mixed ith various minerals substances
like sand, soil,calcium oxalate,chalk powder or other drugs with different inorganic contents to
improve their appearance .

Example: Aloes – NMT 5 %

Ashoka – NMT 11%
Amla – NMT 7%
Nutmeg – NMT 3%
EXPERIMENT NO: 14 DATE:

AIM : To determine Acid soluble ash value, water soluble ash value of given Sample.

REFERENCE :
1. C.K.Kokate,Practical pharmacognosy, fifth edition , vallabh prakashan, page no. 123,124.
2. Biren Shah, A.K.Seth , Rextbook of pharmacognosy and phytochemistry , second edition , page
no. 119.

ACID INSOLUBLE VALUE:

Definition: The acid insoluble ash is the part of the total ash which is insoluble in diluted
hydrochloric acid.
Procedure: Boil the ash obtained for 5 minutes with 25 ml of dilute hydrochloric acid; collect the
insoluble matter in a Gooch crucible, or on an ashless filter paper, wash with hot water and ignite to
constant weight. Calculate the percentage of acid-insoluble ash with reference to the air dried drug.
Significance: Used for determination of earthy matter present on roots, rhizomes and also on leaves.
Crude drugs carry calcium oxalate crystals the amount may vary depending on environmental
conditions
Example;
Agar – NMT 1%
Amla – NMT 2%
Bael- NMT 1%

WATER SOLUBLE ASH:

Definition: The WATER soluble ash is the part of the total ash which is soluble in water.
Procedure: Boil the ash for 5 minutes with 25 ml of water; collect insoluble matter in
a Gooch crucible or on an ashless filter paper, wash with hot water, and ignite for 15 minutes at a
temprature not exceeding 450º. Substract the weight of the insoluble matter from the weight of the
ash; the difference in weight represents the water soluble ash. Calculate the percentage of water-
soluble ash with reference to the airdried drug.
Significance: water soluble ash is used to detect the presence of material exhausted by water.
Example: Ginger – NMT 1.7%
EXPERIMENT NO: 15 DATE:

AIM: Identification of Fibres and Minerals based on chemical tests.

REFERENCE:

 Practical Pharmacognosy, Dr. K.R. Khandelwal and Dr. Vrunda K. Sethi, 24th edition, Nirali
Prakashan, pg. no. 27.1, 27.11, 27.17, 27.20.
 Practical Pharmacognosy, C.K. Kokate, 5th edition, Vallabh Prakashan, pg. no. 183-186.

Drugs Biological Chemical Physical Identification Uses

Source Constituents Characteristics Tests

Absorbent cotton It consists of Entirely Colour: White -Fibres soak in Surgical

wool epidermal cellulose: with Odour: Odourless iodine water- dressings,
(Absorbent trichomes or moisture. Taste: Tasteless dry-sulphuric filtering
cotton, surgical Size: Cotton fibres
hairs of seeds acid(66% v/v) medium,
cotton, purified are 2.5 to 4.5cm in
of cultivated Purplish blue insulation
cotton) length, and 25 to
species of or bluish material.
35µ in diameter.
Gossypium. green colour.
Extra features: Soft,
The -Dissolves in
fine filament like
trichomes ammonical
hairs, slightly off-
freed from copper oxide
white if sterilized.
fatty matter solution.
and adhering -Insoluble in
impurities, dil. Sodium
bleached and hydroxide
sterilized. solution and
Family: soluble in
Malvaceae. sulphuric
acid(66% v/v).
-No stain with
sodium
picrate-picric
acid solution

Jute Strands of Ligno-cellulose Colour: Yellowish Jute + In

(Gunny) phloem fibres (53%), brown phloroglucinol manufacturin
from the stem hemicelluloses Size: 0.80to 5.0mm and g tows; as a
bark of (20%), lignin in length and 10.25µ hydrochloric filtering and
 Corchorus (10%), in diameter. acid: middle straining
capsularis, moisture (13%) Commercial fibres lamella gives medium.
and other are 1-3 metre in red colour
species of length and 30-40µ in ( lignin)
Corchorus. diameter.
Family:
Tiliaceae

Silk Fibres A protein Colour: Yellow Soluble in In

obtained fibroin which Size: 5 to 25 micron cuoxam, preparation
from the on hydrolysis in diameter and sulphuric of sutures,
cocoons of yields amino about 1200 roeters acid(66%) and sieves and
Bombyx acid glycine in length. conc. ligatures.
mori, the and alanine. Extra features: The Hydrochloric
mulberry silk silk threads are very acid.
worm and fine, solid, smooth.
other species
of Bombyx
and
Antheraea
( order
Lepidoptera)
Family:
Bombycidae.

Wool Wool fibres Sulphur Smooth, elastic, Solution of As filtering

are obtained containing curly, hygroscopic wool in and straining
from the protein known and slippery to caustic soda+ medium, in
fleece of as keratin; touch; have lead acetate preparation
sheep Ovis which is rich in tendency to cling solution→ a of dressings
aries (order amino acid together.Solubility: black like domette
Ungulata). cystine. Insoluble in 66% precipitate and crepe
Family: sulphuric acid, conc. (sulphur) bandages and
Bovidae. Hydrochloric acid flannel.
and cuoxam.
OBSERVATION & INFERENCE
 EXPERIMENT NO: 16 DATE:

AIM: To identify unorganized drug by Chemical tests.

REFERENCE:

1. Textbook of Pharmacognosy by C.K. Koakate, S.B. Gokhale, Nirali Prakashan, 43rd edition,
pg no. 34-36
2. Textbook of Pharmacognosy and Phytochemistry by Vinod Rangari, pg no.

THEORY:

Unorganized drugs:

These are derived from plants or animals by some process of extraction and followed by purification,
if necessary.

E.g. Juices, extracts, resins etc.

These are solid, semisolid or liquid in nature.

E.g. Oils, gums and balsams.

Few unorganized crude drugs are commonly describes by their physical characters only. The
description is as below:

1. Gums and mucilages-

Gums are translucent and amorphous substance produced by plants. Gums are usually pathological
products and are produced when the plants is growing under unfavorable conditions. Thus they are
abnormal products of plants metabolism. The gums are produced by the process known as
‘gummosis. Mucilages are also plants products. Similar to gums are regarded to be normal products
of plant metabolism. Mucilages are produced inside the cells of the plant. Mucilages form slimy
masses with water but do not dissolve. mucilages are esters of sulphuric acid, where in ester group is
a polysaccharide complex.
2. Resins and resin combinations-
The resins are of two types:
(i) Synthetic resins
(ii) Natural resins
Natural resins:

The non uniform physical and chemical nature of these substances makes them difficult to define.
They are obtained from plant as well as from animal sources. The resins of animals sources is shellac
or lac which finds number of applications in pharmaceutical industry.

Oleo resins:

When the natural plants resins are accompanied with volatile oils in homogeneous form they are
known as oleo resins. Canada balsam and capaiba are suitable examples of oleo resins.

Balsams:

Aromatic resinous substances of plant origin containing balsamic acids are known as balsams.
Neither Canada balsam nor balsam of Capaiba contains any balsamic acids and hence, they are not
balsams in real sense. The examples are balsam of tolu, benzoin, storax and peru.

Oleo gum resins:

These are the combinations of volatile oils, gums and resins. Sometimes they also contain ether
substance like enzyme e.g. Myrrh and asafetida.

Dried juices:

Theses juices are obtained from fleshy leaves (aloes) of from stems of trees. In all cases incisions are
made to respective part of plants and juice coming out is collected and dried.

Latices:

The latex is product contained in special secretory tissues of certain plants. It is usually a white
aqueous suspension wherein microscopically small particles of oily globules are suspended. These
natural suspension of milky consistency may contain proteins, sugars, minerals and alkaloidal salt in
the true solution, whereas gums, starch and resins I n the suspended form.

Extracts:

The extracts covered under crude drugs differ from galenical extracts. The extract of pharmacognostic
origin consists of extracting the parts of the plant with water followed by concentration, while
pharmaceutical preparations known as extracts are prepared by using alcoholic solutions and
adjusting the products is as standard strength.
DRUGS BIOLOGICAL PHYSICAL CHEMICAL IDENTIFICATION USES
SOURCE CHARACTERISTICS CONSTITUENTS TESTS

AGAR Dried Strips: colorless, Carbohydrate: -Boil agar with Bulk laxative,
(Agar-agar, gelatinous slender, translucent, Polysaccharides water→ forms stiff pharmaceutical aid,
Japanese substance, lustrous, 4 mm wide i.e. agarose and jelly on cooling. in the preparation of
Isinglass) obtained from Bands: yellowish, 4 agaropectin. culture media.
Gelidium cm wide. -Agar solution +
amansii, G. Sheets: 45-60cm long ruthenium red →
cartilagineum, and 10-15 cm wide. pink color.
Flakes or course
G. Pristodes,
powder: grayish -Agar solution (hot)
Gracilaria
white, odorless + BaCl2 reagent→
confervoides,
Taste: mucilaginous white ppt.
P. capillacea Solubility: practically
and other insoluble in cold -Agar solution +
closely allied water, but swells to a Fehling’s solutions
members of gelatinous mass. + heat→ red ppt.
family Soluble in boiling -Agar + Iodine
Rhodophyceae. water. solution→ crimson
to brown color.

-Agar ash (on slide)

+2 drops HCl→
sponge spicules of
diatoms are
observed under
microscope.

-All chemical tests,

mentioned for
gelatin are not
positive.

DRUGS BIOLOGICAL PHYSICAL CHEMICAL IDENTIFICATION

SOURCE CHARACTERISTICS CONSTITUENTS TESTS
GUAR GUM The powder of the Color: yellowish-white Carbohydrates: -Weak iodine
(Guar flour, Jaguar endosperm of the seeds powder. water soluble: solution→ no olive-
gum) of Cyamopsis odor: characteristic guaran (85%) green color.
Taste: gummy proteins (5.7%)
tetragonolobus Linn.
Guar gum -With 2% solution of
 Family: swells rapidly lead acetate→ ppt.
Leguminosae. in water witch
translucent -With Ruthenium
suspension. red→ no pink color.
0.5% Aqueous
-0.1% drug in 20 ml
solution of
water, shake, add 0.5
gum is neutral
ml of hydrogen
to litmus.
peroxide solution,
and 0.5 ml of 0.1%
solution of benzidine
in alcohol: no blue
color.

HONEY A sugar secretion Color: Pale yellow Carbohydrates: Fehling solution

(Madhu, Honey deposited in honey or yellowish brown. Glucose (35%), test→ red ppt.
purified, Mel) comb by the bee, Odor: Characteristic, Fructose (45%),
Apis dorsata, and pleasant. Sucrose (2%), Maltose, Fiehe’s test is not
other species of Apis Taste: Sweet and gum, enzyme invertase, positive (it is for
e.g. A. Indica, A. faintly acid. diastage, inulase. artificial invert
florae, etc. Extra features: sugar)
Family: Apidae. Syrup thick liquid
translucent when
fresh, then opaque
and granular due to
the crystallization of
glucose.

DRUGS BIOLOGICAL PHYSICAL CHEMICAL IDENTIFICATION

SOURCE CHARACTERISTICS CONSTITUENTS TESTS
INDIAN GUM The dried, gummy Color: Tears are Carbohydrates: -Aqueous solution
(Acacia, Babul or exudation from the cream brown to red Arabin, Arabic acid, with lead subacetate
Kikar Gond, Gum stem and branches of in color, powder is enzyme oxidase and solution→
Arabic. Acacia arabica light brown in color. peroxidase. gelatinized form.
Wild. Odor: odorless -With Ruthenium red
Family: Taste: bland and solution→ no pink
Leguminosae. mucilaginous color.
Size and shape: -Aqueous solution of
irregular brown tears gum+ 0.5 ml of
of varying size. solution of
Extra features: tears hydrogen peroxide
with minute fissures, + 0.5 l of solution
brittle in nature, of benzidine in
glossy and alcohol (1%) →
occasionally blue color (oxidase
iridescent.
 enzyme).
-Aqueous solution of
gum + dil. HCl,
boil, Fehling’s
solution A and B,
heat→ red ppt.
(reducing sugars)
-Aq. Solution + borax
→ translucent
mass.
-No blue/ black
coloration with
N/10 Iodine
solution.

-No blue/ black

coloration with 0.1%
FeCl3 solution.

DRUGS BIOLOGICAL PHYSICAL CHEMICAL IDENTIFICATION

SOURCE CHARACTERISTICS CONSTITUENTS TESTS
TRAGACANTH Dried gummy Color: white or pale Carbohydrates: Water -Boil with freshly
(gum tragacanth) exudation obtained yellowish-white soluble part- prepared 10% aq.
by incision from flakes. tragacanthin (8-10%), Ferric chloride
stems and branches Odor: odorless. water insoluble part- solution→ deep
of Astragalus Taste: tasteless bassorin (60-70%) yellow ppt.
Shape: Thin flattered
gummifer Labill and
ribbon like flakes. -Dissolve
other species of
Size: Flakes are 25 x tragacanth and ppt.
Astragalus.
12 x 2 mm copper oxide in
Family:
Solubility: Partly
Leguminosae. conc. Ammonium
soluble in water, hydroxide→ a
swells, insoluble in stringy precipitate.
alcohol.
Extra features: The -Fehling’s solution
gum is horny, test→ red ppt.
translucent, and
transverse with -Drug solution +
longitudinal ridges. lead acetate→
Fracture is short. white ppt.

-Powder + 5%
aqueous caustic
potash→ canary
yellow color.
EXPERIMENT NO: 16 DATE:

AIM: To study Morphology of Crude Drugs.

DRUGS Common Biological Source Chemical Uses

Names Constituents

1] SENNA Senna, Sona Obtained from leaf and pods Glycosides, Laxative, Supply
mukhi of Cassia angustifodia(fam- Antrancene- in habitual/
Leguminosae) Flavonides constipation

2] BEAL Bel Obtained from fruit of Aegle Tannis, Chronic diarrhoe

marelos L. corr(family- Reducing sugar. and Dysentry,
Rutaceace) cooling effect

3] CASSIA Chinese Obtained from dried Volatile oil, Flavouring agent,

cinnamon cinnamomum Cassa stem cinnamic mild astringent,
bark(fam-Lauraceace) aldehyde, etc germicidal

4] AGAR Agar-agar, Obtained from Gelidium Carbohydrate, Bulk laxative

STRIP Japenese, and Pterocladia. polysaccharides nutrient media
lsinglass. (fam.Gracilaria-ceae) preparation,
A] Agarose vaginal capsule
B] Agaropectin and suppositories

5] Dalchini, Obtained from dried bark of Volatile oil, Carminative,

CINNAMON Ceylon taj, shoots of coppocied tree of phenol, Flavouring agent,
BARK etc cinnamomum zeylanicum hydrocarbons. mild astringent.
 Blume(fam-Lauraceae)

6] ISAPGOL Isphapgol Obtained from dried seeds Alkaloids, fixed To relieve gout,
of plantago ovate(fam- oils Epidermal cancer,
Isabghol plantagin-aceae Leukamia

7] Rhizoma Dried rihizome of rheum Glycosides, Laxative

RHUBARB emodiwalls. R.webbianum Tannis, fatty
Resochini oils etc. Antidiarrhoe
(fam. Polygonaceae)
8] Meadow saffron Calcium corm Alkaloids Colchicine, To relieve
COLCHICUM corm, etc and seed are Colchicine gout,
dried corm and Epidermal
strip seed of cancer
Colchicum
Luteumbaker

(fam. Liliceae)

9] Guduchi, Galo, It consist of Clerodane Antipyretic,

TINOSPORA Amrita dried stem of flarnoditerpenec. Analgesic,
Amritavallari Tinospora Antidiabetic
cordifolia wild, Alkaloids.
fam. Anti-
Sequiterpene glucoside inflammatory
Menispermaceae
Phenylpropane.dissachride

10] SQUILL Maritime squill: It consist of Flavonides Used as a hair

BULB Urginea Indica dried bulb of tonic to treat
Urginea Antifungal dandruff
maritime Baker

(fam. Liliaceae)

11] POTATO Marphy, spud, It is dried tuber Carbohydrates Provide

TUBER potato of solanum essential
Minerals nutrients, Vit
(fam. Salanacae) C source

12] ACACIA Ben-Hin Babul, Acacia is the Carbohydrates, Demulcent

dried exudation Polysaccharides, Arabin.
Mal.Karuvelam, from the stem Emulsifying
Babula. and branches of agent, Binding
Acacia Senegal agent.
wild(fam.
Leguminousae)